Pharmaceutical composition containing silibinin and pueraria root extract

ABSTRACT

Disclosed are a pharmaceutical composition for treating non-alcoholic fatty liver and a method for preparing same. The composition is prepared from the following hulk drugs by weight ratio: 8.75-60 parts of silibinin, 15-65 parts of phospholipid, 25-200 parts of Pu&#39;er tea extract, and 5-50 parts of radix puerariae extract.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is the National Stage of International Application No.PCT/CN2016/077038 filed Mar. 22, 2016, which claims the benefit ofChinese application number 201510128638.8, filed Mar. 23, 2015 thedisclosures of which are incorporated herein by reference in theirentireties.

TECHNICAL FIELD

The present invention relates to the technical field of medicines, andparticularly to a pharmaceutical composition containing silibinin farthe treatment of hepatopathy.

BACKGROUND ART

In the late 1960s and 1980s, the pharmaceutists of West Germany withH.wagner as representative extracted the active ingredient from thefruit of the Silybummarianum, which is named as silymarin, a new classof flavonoid having a C-9 substituents, i.e., a flavonoid lignanscondensed with a dihydroflavonol and a phenylpropanoid derivative.Silibinin (silibinin) is one of the main components of silymarin.Pharmacological and toxicological studies have shown that silibinin hasthe effects of protecting and stabilizing the hepatocyte membrane,promoting the recovery of hepatocyte and improving the liver function.Silibinin has different levels of protection and treatment effects onvarious types of hepatic injury caused by hepatic poisons such as carbontetrachloride, thioacetamide, hydroxycholine, phalloidine, mucronatine,etc. And silibinin can be used for treating acute and chronic hepatitis,early hepatocirrhosis, fatty liver, toxic or drug-induced hepatopathy.

The silibinin is poor in water solubility and common organic solvents,resulting in low bioavailability and thereby affecting the clinicalefficacy. To improve the bioavailability thereof, domestic and externalpharmacy workers have made substantial amounts of work. The measures toimprove the absorption of poorly soluble drugs are typically superfinegrinding salinization, and the addition of cosolvent, etc. In recentyears, the studies have shown that the dissolution and bioavailabilityare greatly improved by the methods of formulating into cyclodextrininclusion compound, solid dispersion, synthetic phospholipid complex andformulating into different dosage forms.

From the perspective of solid preparation, the phospholipid complex is amore specific solid dispersion, which has a fixed melting point, is amolecular compound (complex) whose chemical nature is more stable anddifferent from the compound of drug and phospholipid, such compoundsvaries with the types of phospholipid and ratios of drug tophospholipid, and a phospholipid molecule can be bound with a differentnumber of drug molecules. Deduced from the spectroscopy characteristicsof the complex, the drug has a strong interaction with the polar groupsof the phospholipid, which inhibits the free rotation of the singlechains in the molecule, whereas the two long fatty acid chains of thephospholipid do not participate in the complex reaction and are free toshift and wrap the polar portions of the phospholipid to form alipophilic surface, so that the complex shows strong lipid solubility.The complex changes the physiochemical properties of drug, and thusincreases the lipid solubility of the drugs and reduces the watersolubility of the drugs, and promotes the combination of drug moleculesand cell membranes to improve the absorption and increases thebioavailability of the drug.

Pu'er tea is a unique and famous tea in Yunnan province. The localityhas moderate climate, abundant rainfall and is mist-shrouded. Pu'er teais divided into two series by Yunan big leaf species sun-dry tea andreprocessing thereof: the unzymic Pu'er tea by directly re-processinginto the finished product and the enzymic Pu'er tea by re-processingafter the artificial accelerated fermentation, and the patterns of whichare divided into loose tea and compressed tea; natural aging process isalso persistently carried out after the finished products, with theunique qualities gets better.

Pu'er tea is the only post-fermented tea, and substances harmful to thehuman body such as theophylline, tea polyphenols are degraded in thelong process of fermentation, so the product is mild, does not stimulatethe body, and also can promote metabolism, accelerate the digestion andtransformation of fats and toxins in the body. For the problems ofobesity and three-hypes which are puzzling urbanites, Pu'er tea can playa good mitigation effect, such as expelling of toxin, nourishing thestomach, anti-inflammatory, reducing the cholesterol, off lipid andremoving grease, cosmetic slimming. Modern technologies show that Pu'ertea can improve insulin resistance, regulate levels of blood lipid andleptin, etc., and can block the fat accumulation of hepatic parenchymalcell caused by insulin resistance to some extent.

Non-alcoholic fatty liver disease (NAFLD) is a metabolic stress-inducedhepatic injury that is closely related to insulin resistance and geneticsusceptibility, the pathological changes of which are similar toalcoholic fatty liver disease. NAFLD is a clinicopathological syndromecharacterized by steatosis and fat storage of hepatocytes in the hepaticlobule but without history of alcohol abuse. NAFLD shows differentdegrees of hepatic lesion, from simple fatty liver without anyinflammation to severe inflammatory response of severe fibrosis and evencirrhosis, mainly includes 3 types: simple fatty liver, steatohepatitis,fatty cirrhosis.

Non-alcoholic Fatty Liver Disease Treatment

-   1. Prevention of protopathies or associated risk factors.-   2. Basal treatment: developing a reasonable energy intake and diet    adjustment, taking moderate aerobic exercises, correcting bad    lifestyles and behaviors.-   3. Avoiding aggravating hepatic injury: preventing a sharp decline    in weight, drug abuse and other factors that may induce exacerbation    of hepatopathy.-   4. Weight loss: requiring all NAFLD patients who are overweight, and    have visceral obesity and rapid weight gain in the short term to    change the lifestyles to control weight and reduce waist    circumference. Basal treatment for 6 months, weight loss <0.45 kg    per month, or body mass index (BMI) >27 kg/m² combined with blood    lipid, blood glucose, blood pressure and other indicators of more    than two abnormalities may consider adding sibutramine or orlistat    and other obesity drugs, weight loss per week should not exceed 1.2    Kg (children do not exceed 0.5 Kg per week); BMI >40 kg/m² or    BMI >35 kg/m² combined with sleep apnea syndrome and other    obesity-related diseases, may consider the proximal end gastric    bypass procedures to lose weight (II-1, II-2, II-3, III).-   5. Insulin sensitizer, combined with type 2 diabetes, impaired    glucose tolerance, fasting plasma glucose and visceral obesity, may    consider the application of metformin and thiazolidinediones in    order to improve insulin resistance and control of blood glucose    (II-1, II-2, II-3).-   6. Hypolipidemic agents: dyslipidemia, with basal treatment and (or)    application of weight loss and hypoglycemic drugs for more than 3-6    months, is still mixed with hyperlipidemia or hyperlipidemia,    combined with more than 2 risk factors, should consider adding the    use of fibrates, statins or probucol and other hypolipidemic drugs    (II-1, II-2, II-3).-   7. Drugs for hepatopathy: NAFLD associated with hepatic dysfunction,    metabolic syndrome, 3-6 months after basal treatment remains    ineffective, and liver biopsy shows NASH and chronic progression of    the course of the disease, the drug auxiliary treatment for    hepatopathy can be used with antioxidant, anti-inflammatory,    anti-fibrosis, and related drugs (II-1, II-2, II-3, III) such as    polyene phosphatidylcholine, vitamin E, silymarin and    ursodeoxycholic acid can be rationally chosen according to drug    performance, disease activity and stage of the disease, but    multi-drugs should not be applied simultaneously.-   8. Liver transplantation: mainly for NASH-related end-stage    hepatopathy and some cryptogenic J hepatocirrhosis, and the    metabolic condition (III) should be screened before liver    transplantation. BMI >40 kg/m² is contraindication to liver    transplantation (III).

The above treatments have not been used by being mixed together, such asa combination of hypoglycemic and hepatopathy drugs, or a combination oflipid-lowering and hepatopathy drugs. Therefore, the search for apharmaceutical with a variety of health-promoting functions cannot wait.

SUMMARY OF THE INVENTION

In order to solve the above technical problems, the present inventionprovides a pharmaceutical composition and a preparation thereof which istherapeutically effective for non-alcoholic fatty liver diseases, andalso has a synergistic effect on the treatment of non-alcoholic fattyliver diseases when the four are combined in use.

The present invention provides the methods for preparing thepharmaceutical composition and the preparation thereof.

The present invention is achieved by the following technical solutions:

A pharmaceutical composition is prepared from the following bulk drugsby weight ratio:

-   -   8.75-60 parts of silibinin,    -   15-65 parts of phospholipid,    -   25-200 parts of Pu'er tea extract, and    -   5-50 parts of radix puerariae extract.

Preferably, the pharmaceutical composition is prepared from thefollowing bulk drugs by weight ratio:

-   -   25-40 parts of silibinin,    -   30-50 parts of phospholipid,    -   80-120 parts of Pu'er tea extract, and    -   10-35 parts of radix puerariae extract.

Most preferably, the pharmaceutical composition is prepared from thefollowing bulk drugs by weight ratio:

-   -   35 parts of silibinin,    -   42 parts of phospholipid,    -   100 parts of Pu'er tea extract, and    -   20 parts of radix puerariae extract.

The phospholipid of the present invention is a phospholipid or lecithin,which is mainly composed of phosphatidylcholine, preferably soybeanphospholipid.

The role of the phospholipid in the present invention is to promote thedissolution and absorption of pharmaceuticals, silibinin is apharmaceutical with low solubility and low permeability, and thephospholipid is combined therewith to form a phospholipid complex so asto improve solubility of the silibinin, thereby improving thebioavailability of the pharmaceuticals.

Described silibinin and phospholipid are both known from the prior artor commercially available. In order to better exert the efficacy of thepresent invention, the silibinin of the present invention is preferablyprepared by dissolving silymarin in 80% ethanol, filtering and washingthe precipitate with 95% ethanol for three times, collecting theprecipitate. The precipitate is dissolved in anhydrous ethanol,filtered, and the filtrate is added with a certain amount of water toseparate out the precipitate, and the precipitate is collected byfiltration, dried under reduced pressure, pulverized and mixed.

The Pu'er tea extract is commercially available, preferably a DEEPURE®Pu'er tea essence. It is also possible to be prepared according to theprior art. In order to better exert the efficacy of the presentinvention, Pu'er tea essence or Pu'er tea extract is preferably preparedaccording to the method of patents (publication No. CN101961061A,CN101961061B, CN101961425A, CN101961425B, CN101961060A, CN101961059A,CN101961059B).

For example, said Pu'er tea essence is prepared as follows:

-   Step 1, Pu'er tea leaves are decocted with 6-12 times the volume of    water for 2-4 times, 0.5-2 hours each time; extract solution is    filtered, and filtrate is concentrated under reduced pressure and    the temperature of ≤70° C. to the weight of tea leaves:the volume of    concentrate=1:2-1:3;-   Step 2, the concentrate is centrifuged with a centrifuge, the    centrifugate is concentrated under reduced pressure to density of    1.1-1.25 at 45-65° C., concentrated cream is spray dried or    microwave dried to obtain the final product.

Preferably, the steps are present as follows:

-   Step 1, Pu'er tea leaves are decocted with 6-12 times the volume of    vigorously boiling water for 3 times, 0.5-2 hours each time; extract    solution is filtered, and filtrate is concentrated under reduced    pressure and the temperature of ≤70° C. to the weight of tea    leaves:the volume of concentrate=1:2-1:3;-   Step 2, the concentrate is centrifuged with a tripod pendulum type    batch centrifugal, the tripod pendulum is centrifuged with a    tubular-bowl centrifuge, and the centrifugate is concentrated under    reduced pressure to density of 1.1-125 at 45-65° C., concentrated    cream is spray dried or microwave dried to obtain the final product;    wherein tubular-bowl centrifuge condition is: centrifuge speed:    15000-19000 rpm/min; spray drying conditions are: inlet temperature:    140-190° C., outlet temperature: 75-95° C.

Most preferably, the steps are present as follows:

Pu'er tea leaves are decocted with vigorously boiling water for 3 times,the first time decocted for 1.5 h, 10 times the volume of water added;the second time decocted for 1.5 h, 8 times the volume of water added;the third time decocted for 1 h, 8 times the volume of water added,extract solution is filtered, and filtrate is concentrated under reducedpressure and the temperature of ≤70° C. to the weight of tea leaves:thevolume of concentrate=1:2-1:3, the concentrate is centrifuged with atripod pendulum type batch centrifugal, the tripod pendulum iscentrifuged with a tubular-bowl centrifuge, and the centrifugate isconcentrated under reduced pressure to density of 1.1-1.25 at 45-65° C.,concentrated cream is spray dried or microwave dried to obtain the finalproduct;

wherein tubular-bowl centrifuge condition is: centrifuge speed:15000-19000 rpm/min; spray drying conditions are: inlet temperature:140-190° C., outlet temperature: 75-95° C.

The radix puerariae extract is commercially available for dietsupplement or for pharmaceutical use, characterized in that the puerarincontent is not less than 80%.

The above compositions are made by weight ratios, and may be increasedor reduced according to corresponding proportion in productionprocesses, such as large-scale production can be in unit of kg or T(ton); small scale preparations can also be in unit of g. The weight canbe increased or reduced, but the proportions of the weight ratio of bulkdrugs between the components remain unchanged.

The proportions of the above weight ratio are obtained throughscientific screening, for special patients, such as patients with severeor mild symptom, obese or thin patients, the proportions of the amountof composition can be accordingly adjusted, increased or decreased nomore than 10%, the efficacy is substantially constant.

Any pharmaceutically acceptable dosage forms can be formulated in theformulation of a pharmaceutical preparation, the dosage forms areselected from: tablet, sugar coated tablet, film coated tablet, entericcoated tablet, capsule, hard capsule, soft capsule, oral liquid, oralagent, granule, pill, powder, paste, sublimed preparation, suspensionagent, solution, injection, suppository, ointment, emplastrum, creme,spray, patch. Preferably oral preparations, and optimal preferablytablet, capsule, granule.

Some pharmaceutically acceptable carriers can be added into thecompositions of the present invention as needed, the pharmaceuticalpreparations can be prepared using galenic pharmacy conventionaltechniques, such as mixing the pharmaceutically active substances withpharmaceutically acceptable carriers. The pharmaceutically acceptablecarriers are selected from: mannitol, sorbitol, sorbic acid or sylvite,sodium metabisulfite, sodium bisulfite, sodium thiosulfate, cysteinehydrochloride, mercaptoacetic acid, methionine, vitamin A, vitamin C,vitamin E, vitamin D, azone, disodium EDTA, calcium disodium EDTA, thecarbonate, acetate, phosphate of monovalence alkali metal or aqueoussolution thereof hydrochloric acid, acetic acid, sulfuric acid,phosphoric acid, amino acid, sodium chloride, potassium chloride, sodiumlactate, xylitol, maltose, glucose, fructose, dextran, glycine, starch,sucrose, lactose, mannitol, silicon derivative, cellulose and derivatethereof, alginate, gelatin, polyvinyl pyrrolidone, glycerine, propyleneglycol, ethanol, Tween 60-80, Span-80, beeswax, lanolin, liquidparaffin, cetyl alcohol, gallic acid esters, agar, triethanolamine,basic amino acid, urea, allantoin, calcium carbonate, calciumbicarbonate, surfactant, polyethylene glycol, cyclodextrin,beta-cyclodextrin, phospholipid material, kaolin, talc, calciumstearate, magnesium stearate, etc. Preferably, the carrier is one ormore of microcrystalline cellulose, lactose, starch, sodiumcarboxymethylcellulose, low substituted hydroxypropyl cellulose, talc.

When the composition of the present invention is prepared intomedicament, the unit dosage of the medicament may contain 0.1-1,000 mgof the pharmaceutically active substance of the present invention, andthe remainders are pharmaceutically acceptable carriers. Thepharmaceutically acceptable carriers may be 0-99.9% of the totalpreparation weight by weight. Preferably, the pharmaceuticallyacceptable carriers may be 40-70% of the total preparation weight byweight. The usage and dosage of the Chinese traditional medicinecompositions or preparations of the present invention are determinedaccording to the conditions of patients while being used.

The cumulative dissolution rate of the pharmaceutical compositionpreparations according to the present invention, such as tablets,capsules, granules and so on, is not less than 80% while dissolution invitro for 2 h, in a dissolution condition: slurry method, rotation speedof 100 rpm and temperature of 37□, and release medium is: 1000 ml ofhydrochloric acid solution at pH1.2, dosage: 1 capsule/1 tablet/1 baggranules.

A preparation method of the pharmaceutical composition of the presentinvention comprises the following steps:

-   {circle around (1)} taking a prescription amount of raw materials    for later use;-   {circle around (2)} preparation of silibinin complex liquid:    weighing a prescription amount of silibinin and phospholipid, and    dissolving them in the anhydrous ethanol, heating and refluxing to    clarify the solution and continuing to heat for a certain time, then    concentrating the clear solution under reduced pressure to a certain    volume, to obtain the silibinin complex liquid for later use;-   {circle around (2)} granulation: weighing a prescription amount of    Pu'er tea extract as a base material, taking the silibinin complex    liquid prepared in step {circle around (2)} as a feed liquid, and    preparing granules by a fluidization spray method with a fluidized    bed, and drying after the liquid complex is all sprayed in for later    use;-   {circle around (4)} total blending: mixing a prescription amount of    radix puerariae extract with the granules of step {circle around    (3)} uniformly in an equal incremental manner to obtain the    pharmaceutical composition.

The present invention also includes a preparation step {circle around(5)}, taking the pharmaceutical composition of step {circle around (4)}and pharmaceutically acceptable carriers, and preparing pharmaceuticallyacceptable dosage forms according to the conventional preparationprocess.

Further preferably, the preparation method of the pharmaceuticalcomposition of the present invention, comprises the following steps:

-   {circle around (1)} taking a prescription amount of raw materials    for later use;-   {circle around (2)} preparation of silibinin complex liquid:    weighing a prescription amount of silibinin and phospholipid, and    dissolving them in the anhydrous ethanol, heating and refluxing to    clarify the solution and continuing to heat for a certain time, then    concentrating the clear solution under reduced pressure to a certain    volume, to obtain the silibinin complex liquid for later use;-   {circle around (3)} granulation: weighing a prescription amount of    Pu'er tea extract as a base material, taking the silibinin complex    liquid prepared in step {circle around (2)} as a feed liquid, and    preparing granules by a fluidization spray method with a fluidized    bed, and drying after the liquid complex is all sprayed in for later    use;-   {circle around (4)} total blending: mixing a prescription amount of    radix puerariae extract with the granules of step {circle around    (3)} uniformly in an equal incremental manner to obtain the    pharmaceutical composition.-   {circle around (5)} preparation: taking the pharmaceutical    composition and pharmaceutically acceptable carriers to prepare    pharmaceutically acceptable dosage forms.

Wherein the heating time described in step {circle around (2)} is0.5-1.5 hours; the concentrated volume is 5%-20% of the original volumeand the temperature of concentration under reduced pressure is 60-80° C.

Wherein the parameters of the fluidized bed in step {circle around (3)}are: the temperature of the materials is 40-65° C., during thegranulation process, the parameters such as fan frequency, inlet airtemperature and infusion frequency are adjusted to keep the materials ingood fluidization state.

After completing the granulation, the granules are dried for 10-60minutes, and the drying temperature is 55-65° C.

Pu'er tea can improve insulin resistance, regulate the levels of bloodlipid and leptin and other effects, can block the fat accumulation ofhepatic parenchymal cell caused by insulin resistance to a certainextent, combined with the strong free radical scavenging andanti-oxidative stress ability of silibinin, the two has preferableanti-NAFLD (non-alcohol fatty liver) effect.

Ve is added to further enhance the beauty-improving effect of theproduct, and L-carnitine is added to enhance the lipid-lowering andweight-controlling effects of the product.

Hereinafter, the advantageous effects of the present invention will bedescribed by experimental examples.

Experimental Example 1 Dissolution Experiment In Vitro

The dissolutions of the silibinin-phospholipid-Pu'er tea-radix puerariaeextract compositions obtained in embodiments 16-20 are determined underthe following conditions: the selection of the dissolution methods isbased on the properties of the main component silibinin in thecompositions, silibinin, as a medicament with low solubility and lowpermeability, is the fourth category in the BiopharmaceuticsClassification System (BCS), dissolution and absorption thereof are boththe rate-limiting steps, either of which should be resolved at the sametime in order to improve the bioavailability of the pharmaceuticals.Dissolution stage of the silibinin is mainly carried out in the stomach,the absorption stage is mainly carried out in the small intestine, andmeasuring the in-vitro dissolution of the pharmaceutical helps toimprove the bioavailability of the pharmaceuticals. Therefore, thefollowing dissolution method is chosen to evaluate the composition:slurry method, rotation speed of 100 rpm and temperature of 37° C., andrelease medium is: 1,000 ml of hydrochloric acid solution at pH1.2,dosage: 1 capsule/1 tablet/1 bag granules. The sampling points are: 15,30, 45, 60, 90, 120 min. The cumulative dissolution is determined. Theresults are shown in Table 1 below.

TABLE 1 Cumulative dissolubility (%) Time Embodiment EmbodimentEmbodiment Embodiment Embodiment Embodiment Embodiment Embodiment (min)1 16 17 18 18 20 22 23 15 18.90% 14.45% 8.98% 15.06% 9.74% 7.11% 7.83%7.08% 30 41.28% 39.22% 29.40% 55.75% 24.11% 36.87% 20.19% 26.11% 4569.83% 71.98% 37.67% 66.24% 51.98% 58.33% 49.02% 54.26% 60 79.42% 78.67%72.90% 69.51% 70.34% 75.42% 71.90% 78.23% 90 83.92% 81.92% 78.43% 80.96%78.87% 79.21% 81.42% 81.38% 120 85.98% 82.35% 81.11% 81.92% 81.38%82.65% 84.38% 86.02%

The dissolution determination of the reference preparation(silibinin-phospholipid complex preparation, laboratory homemade) iscarried out, and compared with the silibinin-phospholipid-Pu'ertea-radix puerariae extract compositions prepared in embodiments 16-20,and the results are as shown in FIG. 1. It can be seen from the data inTable 1 and the graph of FIG. 1:

The in-vitro release of the silibinin-phospholipid-Pu'er tea-radixpuerariae extract composition prepared by the preparation method of thepresent invention is significantly better than that of the referencepreparation silibinin-phospholipid complex, surprisingly, the cumulativedissolution of the composition in the hydrochloric acid solution atpH1.2 for 2 h reaches to more than 80%, nearly completely dissolved,which is doubled as compared with that of the reference preparation,resolving the problems of low solubility and low bioavailability ofsilibinin that have always existed, which will provide the basis for thestudies of dose setting and in-vivo safety and efficacy of silibinincomposition in the future.

Pooling the data of in-vitro dissolution experiments and in-vivopharmacological researches, the present invention further improves thedissolution of the pharmaceutical by combining thesilibinin-phospholipid complex with Pu'er tea and radix puerariaeextract, by continuing the absorption improvements of the drug byincreasing the compatibility of the drug and the biofilm after thecombination of silibinin and phospholipid, from two aspects of improvingthe dissolution and absorption to improve the bioavailability of themain component silibinin.

Experimental Example 2 Evaluation of Pharmacodynamics In Vivo

1 Experimental Animals

80 mice with SPF grade and 6-week-old male C57 BL/6J leptin-deficient(ob/ob), 10 mice with SPF grade and 6 weeks old male C57 BL/6J (ob/m),provided by the Beijing Huafukang Bioscience Co., Inc., raised in TaslyInstitute's pharmacological toxicology research center barrier animalroom, at the temperature of 20° C.-25° C., relative humidity of 60%, 5mice in each cage, lighting time of 12 hours, timely and quantitativefeed, ob/ob mice are fed with high fat diet (HFD, D12492), C57 BL/6Jmice are fed with normal diet, both are provided by Beijing HuafukangBioscience Co., Inc., and free drinking water, daily replacement ofpadding.

2 Tested Substances

Silibinin-phospholipid complex, provided by Tasly Pharmaceutical Co.,Ltd., lot No. 500902031. Pu'er tea extract, brown powder, supplied byTasly Pharmaceutical Co., Ltd., lot No. Z001 PE(2014)C06(H); radixpuerariae extract, provided by Shanxi Yongyuan Bio-Tech Co., Ltd., lotNo. YYP140717; the above tested substances are stored in the samplecabinet of the test room of Pharmacology Institute to be protected fromlight at room temperature.

3 Experimental Methods

3.1 Experimental Dose Design and Grouping

The silibinin-phospholipid complex is administered to the experimentalanimals at a daily dose of 3 g (containing silibinin 420 mg, soybeanphospholipid 504 mg); the Pu'er tea extract is administered to theexperimental animals at a daily dose of 1.2 g. The compatibilityproportions and the experimental dose designs of the five differentcompositions are shown in Table 2, the dose of the experimental animalsis set to the corresponding equivalent dose of the corresponding testedsubstances, the formula for calculation is as follows:animal experimental dose=recommended human dose/60 kg*12.3

TABLE 2 Dose of different compositions Recommended daily dosage (mg)Experiment animal dose design (mg/kg) Prescription Combi- Combi- Combi-Combi- Combi- Combi- Combi- Combi- Combi- Combi- pharmaceuticals nation1 nation 2 nation 3 nation 4 nation 5 nation 1 nation 2 nation 3 nation4 nation 5 Silibinin 420 420 420 105 105 86.10 86.10 86.10 21.53 21.53phospholipid 504 780 780 195 195 103.32 160.00 160.00 40.00 40.00 Pu'ertea extract 1200 1200 2400 300 1200 246.00 246.00 492.00 61.50 246.00Radix puerariae 240 240 60 120 120 49.20 49.20 12.30 24.60 24.60 extract

3.2 Administration of Tested Substances

After 1 week of adaptive feeding, 80 ob/ob mice of 6-week-old arerandomly divided into 8 groups: a model group, a silibinin-phospholipidcomplex group, a Pu'er tea extract group, a combination 1 group, acombination 2 group, a combination 3 group, a combination 4 group, and acombination 5 group, 10 mice for each group. Another 10 C57BL/6J mice of6-week-old are in a normal group. Normal group mice are fed with normaldiet, the model group and the administration group are fed with high fatdiet (HFD, D12492). In addition, the mice in different drug interventiongroups are given the corresponding doses of drugs by means ofintragastric administration, the doses of the five compositions areshown in Table 1, the normal group and the model group are given thesame amount of distilled water, continuous intragastric administrationfor 6 weeks.

The mice are free to eat and drink during the experiment, weekly weight,and the doses are adjusted according to the body weight After the lastadministration, fasting for 12 h, but water is given, weighing the bodyweight, extracting rats' eyeballs to collect blood and then put them todeath by breaking their necks, and the liver is harvested rapidly,physiological saline rinsing, filter paper blotting and preserved in a−20° C. refrigerator after weighing.

3.3 Detecting Indicators and Methods

3.3.1 General Observation

The weights of mice in each group are measured weekly during theexperiment.

3.3.2 Calculation of the Liver Index and Observation of the GeneralMorphology of the Liver

After finishing the experiment, the liver is weighed and the liver indexis calculated, the liver index (%)=liver wet weight/body weight*100%.

3.3.3 Determination of Serum Biochemical Indexes

Blood of all the mice are collected by extracting rats' eyeballs andcentrifuged at 3000 r/min for 15 minutes, the serum is separated andcollected in an EP tube and stored at −20° C. refrigerator for lateruse. The content of glutamic oxaloacetic transaminase (AST),glutamicpyruvic transaminase (ALT), total cholesterol (TC), low-densitylipoprotein cholesterol (LDL-C) in serum are measured by 7020 automaticbiochemistry instrument.

3.3.4 Insulin Resistant Index

Serum FINS is detected using the Elisa kit and the insulin resistanceindex is calculated by the formula.

${{Home} - {IR}} = \frac{{FBG} \times {FINS}}{22.5}$

3.3.5 Liver Histopathological Examination

Frozen sections are prepared from frozen liver tissues and the degree ofhepatic steatosis is observed by oil red O staining. Oil red O stainingoperation steps: frozen slicing→sufficiently washing with distilledwater→staining with oil red O diluent in the dark for 10-15minutes→taking out 6 ml of oil red O saturated liquid, adding 4 ml ofdistilled water, leaving it for 5-10 minutes and filtrating for lateruse→differentiating to interstitial clear under mirror with 60%ethanol→washing with water→nuclear counter staining withhematoxylin→washing with water→sealing piece with neutral gum→microscopeobservation.

3.4 Data Processing

SPSS 15.0 statistical software is used for analysis, the data areexpressed as mean±standard deviation, the t test is used to analyzewhether there's any difference between the two groups before and aftertreatment or not, and the difference is statistically significant withP<0.05.

4 Experimental Results

4.1 the Effects of Each Tested Substance on Body Weight

Mice in each group are measured once a week during the experiment, andthe effects of each group of medicaments on body weight of micesuffering from non-alcoholic fatty liver diseases in each group areobserved. As shown in Table 3, the weight of mice in normal group isincreased slowly and the weight of mice in model group is increased morerapidly. After 6 weeks of administration, the other administrationgroups except for the silibinin-phospholipid complex group could inhibitthe weight increases of mice in different degrees (P<0.05), a combineduse of silibinin-phospholipid complex, Pu'er tea extract and radixpuerariae extract is significantly superior to using silibinin alone.

TABLE 3 Effect of test substances on body weight of mice (g, n = 10, x ±S) Body weight (g) Before Week 2 after Week 4 after Week 6 after Groupadministration administration administration administration Normal 24.36± 1.64 26.64 ± 2.50  27.65 ± 1.54  28.02 ± 2.03  Model 49.88 ± 3.5156.53 ± 2.12  60.15 ± 2.55  62.87 ± 2.75  Silibinin-phospholipid 49.55 ±3.23 55.57 ± 4.13  59.66 ± 5.00  63.10 ± 4.83  complex Combination 150.43 ± 3.72 51.07 ± 2.56** 53.10 ± 4.06** 55.25 ± 4.20** Combination 249.90 ± 3.53 53.87 ± 3.64*  56.56 ± 4.77*  59.74 ± 3.61*  Combination 349.75 ± 4.00 53.82 ± 3.50*  54.80 ± 3.37** 59.64 ± 2.72*  Combination 450.52 ± 3.71 50.98 ± 2.76** 52.01 ± 4.25** 54.16 ± 4.32** Combination 550.88 ± 4.18 51.91 ± 2.83** 53.44 ± 3.56** 55.71 ± 3.88** *compared withthe model group P < 0.05; **compared with the model group P < 0.01;

4.2 the Effects of Each Tested Substance on Liver Index

As shown in Table 4, the body weight, liver wet weight and liver indexof mice in the model group are significantly increased compared with thenormal group (P<0.01), and the silibinin-phospholipid complex candecrease the wet weight of the liver and liver index (P<0.05), but hasno significant effect on body weight of the mice (P>0.05); all the fivecompositions of silibinin-phospholipid complex, Pu'er tea extract andradix puerariae extract can significantly reduce the wet weight andliver index of the liver of mice (P<0.05), the effect of a combined useof the three is significantly better than that of using thesilibinin-phospholipid complex alone.

TABLE 4 Effects of each tested substance on liver index of mice GroupBody weight (g) Liver wet weight (g) Liver mass index % Normal 28.16 ±2.09  1.096 ± 0.106  3.927 ± 0.604  Model 62.77 ± 2.68  4.328 ± 0.297 6.902 ± 0.492  Silibinin-phospholipid 63.66 ± 5.04  3.997 ± 0.376* 6.302 ± 0.670*  complex Combination 1 54.00 ± 2.23** 2.636 ± 0.421**4.843 ± 0.792*  Combination 2 59.29 ± 3.73*  3.462 ± 0.675** 5.922 ±1.171** Combination 3 58.43 ± 3.88** 3.000 ± 0.710** 5.158 ± 1.267**Combination 4 51.07 ± 2.98** 2.436 ± 0.382** 4.775 ± 0.716** Combination5 53.77 ± 2.14** 2.736 ± 0.230** 5.096 ± 0.470** *compared with themodel group P < 0.05; **compared with the model group P < 0.01;

4.3 Effects of Each Tested Substance on Blood Lipid, Liver Function andInsulin Resistance Indexes of Mice

As shown in Table 5, the levels of serum TC, LDL, ALT, AST and insulinresistance index are significantly increased in non-alcoholic fattyliver model mice compared with the normal group (P<0.05); there is nosignificant improvement in the abnormal elevation of thesilibinin-phospholipid complex (P>0.05); the silibinin-phospholipidcomplex, Pu'er tea extract and radix puerariae extract compatibilitygroup of different proportions can significantly reduce TC, LDL-C, ALT,AST and insulin resistance indexes (P<0.05), and the effect is betterthan that of using the silibinin-phospholipid complex alone.

TABLE 5 Effects of each tested substance on blood lipid, liver functionand insulin resistance indexes of mice Group TC LDL-C ALT AST Normal2.99 ± 0.25  0.26 ± 0.03  51.93 ± 20.99  118.62 ± 44.42  Model 10.41 ±1.18   2.33 ± 0.54  425.48 ± 90.03  205.61 ± 22.74 Silibinin-phospholipid 10.35 ± 1.12   2.22 ± 0.32  374.34 ± 56.78 182.61 ± 28.50  complex Combination 1 7.14 ± 0.96** 1.13 ± 0.20** 236.70± 92.42** 123.30 ± 24.39** Combination 2 9.31 ± 1.02*  1.69 ± 0.29* 331.35 ± 98.22*  178.31 ± 29.61*  Combination 3 8.86 ± 1.21*  1.54 ±0.35** 321.69 ± 112.79* 163.28 ± 34.78*  Combination 4 6.08 ± 1.12**1.03 ± 0.25** 181.61 ± 46.79** 126.30 ± 35.62** Combination 5 7.34 ±0.94** 1.15 ± 0.18** 247.81 ± 87.55** 138.30 ± 34.07** *compared withthe model group P < 0.05; **compared with the model group P < 0.01;

4.4 Effects of Each Tested Substance on Liver Pathology of Mice

Oil red O staining: according to the size and number of red particles inhepatocytes of liver frozen issues stained by Oil red O under lightmicroscope, it is divided into mild, moderate and severe type. Mild,that is, ⅓-⅔ of red granules are shown per unit area under lightmicroscope, graded as 1 point; moderate, that is, more than ⅔ of thehepatocytes containing red particles, graded as 2 points; severe, thatis, almost all of the hepatocytes containing red particles, graded as 3points; no steatosis is observed, graded as 0 points.

As shown in Table 6, steatosis occurred in nearly all the hepatocytes inthe liver tissues of the model group, and the pathological scores aresignificantly increased than that in the normal group (P<0.01); there isno significant improvement on liver pathological scores by usingsilibinin-phospholipid complex or Pu'er tea extract alone (P>0.05); thecombination of different proportions of the Pu'er tea extract,silibinin-phospholipid complex and radix puerariae extract cansignificantly improve the liver steatosis, reduce the pathologicalscores (P<0.05), and the effect is better than that of using thesilibinin-phospholipid complex and the Pu'er tea extract alone.

TABLE 6 Effects of each tested substance on liver pathology of mice Oilred O staining Group pathological score Normal 0.000 ± 0.000  Model2.667 ± 0.707  Silibinin-phospholipid 2.444 ± 0.726  complex Pu'er teaextract 2.200 ± 0.919  Combination 1 1.889 ± 0.601* Combination 2 2.000± 0.535* Combination 3 2.000 ± 0.500* Combination 4 1.778 ± 0.833*Combination 5 1.900 ± 0.568* *compared with the model group P < 0.05;**compared with the model group P < 0.01;

5 Experimental Conclusions

The above experimental results show that: the body weight, liver index,blood lipid, ALT, AST and insulin resistance index are significantlyincreased in the mice of the non-alcoholic fatty liver model groupcompared with those in the blank group, and the liver tissues are severesteatosis. Pu'er tea can improve insulin resistance, regulate bloodlipids, radix puerariae has blood lipid-regulating and anti-inflammatoryeffects, and can also improve detoxification function of the liver andrepair damaged liver cells; while silibinin has strong free radicalscavenging and anti-oxidative stress ability, the combination use of thethree has a significant improvement in liver steatosis and preventiveand therapeutic effect on non-alcoholic fatty liver.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is an in-vitro release curve, wherein, each sample is referencepreparation of Shui Lin Jia, Shui Lin Jia without Pu'er tea, andsilibinin-phospholipid-Pu'er tea compositions prepared in embodiments16-20.

DETAILED DESCRIPTION OF THE INVENTION

The present invention is further illustrated by the following specificexamples, but not intended to limit the present invention.

Embodiment 1

Taking 26.25 g of silibinin, 45 g of soybean phospholipid, 75 g of Pu'ertea extract, and 15 g of radix puerariae extract.

{circle around (1)} Preparation of silibinin complex liquid: weighing aprescription amount of silibinin, soybean phospholipid, and dissolvingthem in the anhydrous ethanol, heating and refluxing to clarify thesolution and continuing to heat for 1 h, then concentrated under reducedpressure and recycling the ethanol to 15% of the original volume forlater use;

{circle around (2)} Granulation: weighing a prescription amount of Pu'ertea extract as a base material, taking the silibinin complex liquidprepared in step {circle around (1)} as a feed liquid, preparing thegranules by a fluidization spray method with a fluidized bed,controlling the temperature of materials at 40° C., drying at 60° C. for20 min after the liquid complexes are all sprayed in for later use;

{circle around (3)} Mixing a prescription amount of radix puerariaeextract and the granules prepared in step {circle around (2)} uniformlyin an equal incremental manner, bagging, made into 1,000 bags ofgranules.

Embodiment 2

Taking 180 g of silibinin, 195 g of soybean phospholipid, 600 g of Pu'ertea extract, and 150 g of radix puerariac extract.

{circle around (1)} Preparation of silibinin complex liquid: weighing aprescription amount of silibinin, soybean phospholipid, and dissolvingthem in the anhydrous ethanol, heating and refluxing to clarify thesolution and continuing to heat for 1.5 h, then concentrated underreduced pressure and recycling the ethanol to 20% of the original volumefor later use;

{circle around (2)} Granulation: weighing a prescription amount of Pu'ertea extract as a base material, taking the silibinin composite liquidprepared in step {circle around (1)} as a feed liquid, preparing thegranules by a fluidization spray method with a fluidized bed,controlling the temperature of materials at 65° C., drying at 65° C. for60 min after the liquid complexes are all sprayed in for later use;

{circle around (3)} Mixing a prescription amount of radix puerariacextract and the granules prepared in step {circle around (2)} uniformlyin an equal incremental manner, bagging, made into 1,000 bags ofgranules.

Embodiment 3

Taking 26.25 g of silibinin, 195 g of phospholipid, 600 g of Pu'er teaextract, and 15 g of radix puerariae extract.

{circle around (1)} Preparation of silibinin complex liquid: weighing aprescription amount of silibinin, soybean phospholipid, and dissolvingthem in the anhydrous ethanol, heating and refluxing to clarify thesolution and continuing to heat for 0.5 h, then concentrated underreduced pressure and recycling the ethanol to 5% of the original volumefor later use;

{circle around (2)} Granulation: weighing a prescription amount of Pu'ertea extract as a base material, taking the silibinin composite liquidprepared in step {circle around (1)} as a feed liquid, preparing thegranules by a fluidization spray method with a fluidized bed,controlling the material temperature at 50° C., drying at 55° C. for 10min after the liquid complexes are all sprayed in for later use;

{circle around (3)} Mixing a prescription amount of radix puerariaeextract and the granules prepared in step {circle around (2)} uniformlyin an equal incremental manner, bagging, made into 1,000 bags ofgranules.

Embodiment 4

Taking 26.25 g of silibinin, 195 g of phospholipid, 75 g of Pu'er teaextract and 150 g of radix puerariae extract, and preparing 1,000 bagsof granules according to the method of Embodiment 1.

Embodiment 5

Taking 180 g of silibinin, 45 g of phospholipid, 75 g of Pu'er teaextract and 15 g of radix puerariae extract, and preparing 1,000 bags ofgranules according to the method of Embodiment 1.

Embodiment 6

Taking 180 g of silibinin, 45 g of phospholipid, 600 g of Pu'er teaextract and 150 g of radix puerariae extract, and preparing 1,000 bagsof granules according to the method of Embodiment 1.

Embodiment 7

Taking 180 g of silibinin, 195 g of phospholipid, 75 g of Pu'er teaextract and 150 g of radix puerariae extract, and preparing 1,000 bagsof granules according to the method of Embodiment 1.

Embodiment 8

Taking 26.25 g of silibinin, 48.75 g of phospholipid, 75 g of Pu'er teaextract and 30 g of radix puerariae extract, and preparing 1,000 bags ofgranules according to the method of Embodiment 1.

Embodiment 9

Taking 26.25 g of silibinin, 48.75 g of phospholipid, 300 g of Pu'er teaextract and 30 g of radix puerariae extract, and preparing 1,000 bags ofgranules according to the method of Embodiment 1.

Embodiment 10

Taking 52.5 g of silibinin, 97.5 g of phospholipid, 300 g of Pu'er teaextract and 7.5 g of radix puerariae extract, and preparing 1,000 bagsof granules according to the method of Embodiment 1.

Embodiment 11

Taking 75 g of silibinin, 90 g of phospholipid, 240 g of Pu'er teaextract and 36 g of radix puerariae extract, and preparing 1,000 bags ofgranules according to the method of Embodiment 1.

Embodiment 12 Preparation of the Composition

Taking 90 g of silibinin, 108 g of phospholipid, 270 g of Pu'er teaextract and 90 g of radix puerariae extract, and preparing 1,000 bags ofgranules according to the method of Embodiment 1.

Embodiment 13 Preparation of the Composition

Taking 105 g of silibinin, 126 g of phospholipid, 300 g of Pu'er teaextract and 60 g of radix puerariae extract, and preparing 1,000 bags ofgranules according to the method of Embodiment 1.

Embodiment 14 Preparation of the Composition

Taking 105 g of silibinin, 126 g of phospholipid, 300 g of Pu'er teaextract and 60 g of radix puerariae extract, and preparing 1,000 bags ofgranules according to the method of Embodiment 1.

Embodiment 15 Preparation of the Composition

Taking 120 g of silibinin, 150 g of phospholipid, 360 g of Pu'er teaextract and 105 g of radix puerariae extract, and preparing 1,000 bagsof granules according to the method of Embodiment 1.

Embodiment 16

Taking the granules of Embodiment 8, adding 556 g of microcrystallinecellulose and 64 g of sodium carboxymethyl starch, mixing uniformly,encapsulated into capsules to obtain 1,000 capsules.

Embodiment 17

Taking the granules of Embodiment 9, adding 331 g of microcrystallinecellulose and 64 g of sodium carboxymethyl starch, mixing uniformly,encapsulated into capsules to obtain 1,000 capsules.

Embodiment 18

Taking the granules of Embodiment 10, adding 311 g of microcrystallinecellulose and 32 g of sodium carboxymethyl starch, mixing uniformly,encapsulated into capsules to obtain 1,000 capsules.

Embodiment 19

Taking the granules of Embodiment 13, adding 145 g of microcrystallinecellulose and 64 g of sodium carboxymethyl starch, mixing uniformly,encapsulated into capsules to obtain 1,000 capsules.

Embodiment 20

Taking the granules of Embodiment 14, adding 76 g of microcrystallinecellulose and 64 g of sodium carboxymethyl starch, mixing uniformly,encapsulated into capsules to obtain 1,000 capsules.

Embodiment 21

Taking the composition of Embodiment 8, adding 400 g of lactose, 156 gof starch, and 64 g of sodium carboxymethyl starch, mixing uniformly,encapsulated into capsules to obtain 1,000 capsules.

Embodiment 22

Taking the composition of Embodiment 13, adding 115 g of lactose, 10 gof tale powder, and 84 g of low-substituted hydroxypropyl cellulose,mixing uniformly, encapsulated into capsules to obtain 1,000 capsules.

Embodiment 23

Taking the composition of Embodiment 13, adding 145 g ofmicrocrystalline cellulose and 64 g of sodium carboxymethyl starch,mixing uniformly, encapsulated into No. 0 capsules to obtain 1,000capsules.

The invention claimed is:
 1. A pharmaceutical composition comprising8.75-60 parts by weight of silibinin; 15-65 parts by weight ofphospholipid; 25-200 parts by weight of Pu'er tea extract; and 5-50parts by weight of radix puerariae extract.
 2. The pharmaceuticalcomposition of claim 1 comprising 25-40 parts by weight of silibinin;30-50 parts by weight of phospholipid; 80-120 parts by weight of Pu'ertea extract; and 10-35 parts by weight of radix puerariae extract. 3.The pharmaceutical composition of claim 1 comprising 35 parts by weightof silibinin; 42 parts by weight of phospholipid; 100 parts by weight ofPu'er tea extract; and 20 parts by weight of radix puerariae extract. 4.The pharmaceutical composition of claim 1, wherein puerarin in the radixpuerariae extract is not less than 80%.
 5. A pharmaceutical preparationcomprising the pharmaceutical composition according to claim 1, furthercomprising pharmaceutically acceptable carriers; wherein thepharmaceutically acceptable carriers are 0.1-99.9% of the totalpreparation by weight.
 6. The pharmaceutical preparation according toclaim 5, wherein the pharmaceutically acceptable carriers are selectedfrom the group consisting of sorbitol, sorbic acid or sylvite, sodiummetabisulfite, sodium bisulfite, sodium thiosulfate, cysteinehydrochloride, mercaptoacetic acid, methionine, vitamin A, vitamin C,vitamin E, vitamin D, azone, disodium EDTA, calcium disodium EDTA,hydrochloric acid, acetic acid, sulfuric acid, phosphoric acid, aminoacid, sodium chloride, potassium chloride, sodium lactate, xylitol,maltose, glucose, fructose, dextran, glycine, starch, sucrose, lactose,mannitol, silicon derivative, cellulose and derivate thereof, alginate,gelatin, polyvinyl pyrrolidone, glycerine, propylene glycol, ethanol,polysorbate 60-80, sorbitan monooleate, beeswax, lanolin, liquidparaffin, cetyl alcohol, gallic acid esters, agar, triethanolamine,basic amino acid, urea, allantoin, calcium carbonate, calciumbicarbonate, surfactant, polyethylene glycol, cyclodextrin,beta-cyclodextrin, phospholipid, kaolin, talc, calcium stearate,magnesium stearate, microcrystalline cellulose, and a carbonate,acetate, or phosphate salt of a monovalent alkali metal or aqueoussolution thereof.
 7. The pharmaceutical preparation according to claim5, wherein the pharmaceutical preparation is selected from the groupconsisting of tablet, sugar coated tablet, film coated tablet, entericcoated tablet, capsule, hard capsule, soft capsule, oral liquid, oralagent, granule, pill, powder, paste, sublimed preparation, supensoidagent, solution, injection, suppository, ointment, emplastrum, creme,spray, and patch.
 8. The pharmaceutical composition of claim 2, whereinpuerarin in the radix puerariae extract is not less than 80%.
 9. Apharmaceutical preparation comprising the pharmaceutical compositionaccording to claim 2, further comprising pharmaceutically acceptablecarriers; wherein the pharmaceutically acceptable carriers are 0.1-99.9%of the total preparation by weight.
 10. The pharmaceutical preparationaccording to claim 9, wherein the pharmaceutically acceptable carriersare selected from the group consisting of sorbitol, sorbic acid orsylvite, sodium metabisulfite, sodium bisulfite, sodium thiosulfate,cysteine hydrochloride, mercaptoacetic acid, methionine, vitamin A,vitamin C, vitamin E, vitamin D, azone, disodium EDTA, calcium disodiumEDTA, hydrochloric acid, acetic acid, sulfuric acid, phosphoric acid,amino acid, sodium chloride, potassium chloride, sodium lactate,xylitol, maltose, glucose, fructose, dextran, glycine, starch, sucrose,lactose, mannitol, silicon derivative, cellulose and derivate thereof,alginate, gelatin, polyvinyl, pyrrolidone, glycerine, propylene glycol,ethanol, polysorbate 60-80, sorbitan monooleate, beeswax, lanolin,liquid paraffin, cetyl alcohol, gallic acid esters, agar,triethanolamine, basic amino acid, urea, allantoin, calcium carbonate,calcium bicarbonate, surfactant, polyethylene glycol, cyclodextrin,beta-cyclodextrin, phospholipid, kaolin, talc, calcium stearate,magnesium stearate, microcrystalline cellulose, and a carbonate,acetate, or phosphate salt of a monovalent alkali metal or aqueoussolution thereof.
 11. The pharmaceutical preparation according to claim9, wherein the pharmaceutical preparation is selected from the groupconsisting of tablet, sugar coated tablet, film coated tablet, entericcoated tablet, capsule, hard capsule, soft capsule, oral liquid, oralagent, granule, pill, powder, paste, sublimed preparation, supensoidagent, solution, injection, suppository, ointment, emplastrum, creme,spray, and patch.
 12. The pharmaceutical composition of claim 3, whereinpuerarin in the radix puerariae extract is not less than 80%.
 13. Apharmaceutical preparation comprising the pharmaceutical compositionaccording to claim 3, further comprising pharmaceutically acceptablecarriers; wherein the pharmaceutically acceptable carriers are 0.1-99.9%of the total preparation by weight.
 14. The pharmaceutical preparationaccording to claim 13, wherein the pharmaceutically acceptable carriersare selected from the group consisting of sorbitol, sorbic acid orsylvite, sodium metabisulfite, sodium bisulfite, sodium thiosulfate,cysteine hydrochloride, mercaptoacetic acid, methionine, vitamin A,vitamin C, vitamin E, vitamin D, azone, disodium EDTA, calcium disodiumEDTA, hydrochloric acid, acetic acid, sulfuric acid, phosphoric acid,amino acid, sodium chloride, potassium chloride, sodium lactate,xylitol, maltose, glucose, fructose, dextran, glycine, starch, sucrose,lactose, mannitol, silicon derivative, cellulose and derivate thereof,alginate, gelatin, polyvinyl pyrrolidone, glycerine, propylene glycol,ethanol, polysorbate 60-80, sorbitan monooleate, beeswax, lanolin,liquid paraffin, cetyl alcohol, gallic acid esters, agar,triethanolamine, basic amino acid, urea, allantoin, calcium carbonate,calcium bicarbonate, surfactant, polyethylene glycol, cyclodextrin,beta-cyclodextrin, phospholipid, kaolin, talc, calcium stearate,magnesium stearate, microcrystalline cellulose, and a carbonate,acetate, or phosphate salt of a monovalent alkali metal or aqueoussolution thereof.
 15. The pharmaceutical preparation according to claim13, wherein the pharmaceutical preparation is selected from the groupconsisting of tablet, sugar coated tablet, film coated tablet, entericcoated tablet, capsule, hard capsule, soft capsule, oral liquid, oralagent, granule, pill, powder, paste, sublimed preparation, supensoidagent, solution, injection, suppository, ointment, emplastrum, creme,spray, and patch.
 16. A method of preparing the pharmaceuticalpreparation according to claim 5, comprising: (1) taking a prescriptionamount of raw materials; (2) preparing a silybin complex liquid byweighing a prescription amount of silybin and phospholipid, anddissolving them in the anhydrous ethanol, heating and refluxing toclarify the solution, then concentrating the clear solution underreduced pressure to a concentrated volume, to obtain the silybin complexliquid; (3) granulating by spraying a prescription amount of Pu'er teaextract with the silybin complex liquid by a fluidization spray methodwith a fluidized bed, and drying; (4) blending by mixing radix puerariaeextract and the granules of step (3) uniformly to obtain thepharmaceutical composition; and (5) combining the pharmaceuticalcomposition with pharmaceutically acceptable carriers.
 17. The methodaccording to claim 16, wherein the heating time in step (2) is 0.5-1.5hours; the concentrated volume is 5%-20% of the original volume and thetemperature of the concentration under reduced pressure is 60-80° C.;the parameters of the fluidized bed in step (3) are that the temperatureof the materials is 40-65° C., and adjusting fan frequency, inlet airtemperature and infusion frequency to keep the materials in a goodfluidization state during the granulation process; and wherein after thegranulation is completed, the granules are dried for 10-60 minutes at55-65° C.
 18. A method of treatment, comprising administering atherapeutically effective amount of the pharmaceutical composition ofclaim 1 to a subject in need thereof, wherein the treatment is fornon-alcoholic fatty liver disease, reducing fat, losing weight, orbeautifying skin.
 19. A method of treatment, comprising administering atherapeutically effective amount of the pharmaceutical composition ofclaim 5 to a subject in need thereof, wherein the treatment is fornon-alcoholic fatty liver disease, reducing fat, losing weight, orbeautifying skin.
 20. A method of preparing the pharmaceuticalpreparation according to claim 9, comprising: (1) taking a prescriptionamount of raw materials; (2) preparing a silybin complex liquid byweighing a prescription amount of silybin and phospholipid, anddissolving them in the anhydrous ethanol, heating and refluxing toclarify the solution, then concentrating the clear solution underreduced pressure to a concentrated volume, to obtain the silybin complexliquid; (3) granulating by spraying a prescription amount of Pu'er teaextract with the silybin complex liquid by a fluidization spray methodwith a fluidized bed, and drying; (4) blending by mixing radix puerariaeextract and the granules of step (3) uniformly to obtain thepharmaceutical composition; and (5) combining the pharmaceuticalcomposition with pharmaceutically acceptable carriers; wherein theheating time in step (2) is 0.5-1.5 hours; the concentrated volume is5%-20% of the original volume and the temperature of the concentrationunder reduced pressure is 60-80° C.; the parameters of the fluidized bedin step (3) are that the temperature of the materials is 40-65° C., andadjusting fan frequency, inlet air temperature and infusion frequency tokeep the materials in a good fluidization state during the granulationprocess; and wherein after the granulation is completed, the granulesare dried for 10-60 minutes at 55-65° C.